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本科毕业设计(论文)
英文文献翻译
学院名称:
电气信息工程学院
专业:
电子信息工程
班级:
14电子1B
学号:
201496111
姓名:
李鹏
指导教师姓名:
指导教师职称:
二〇一六年六月
Part1:
cellengineering
In2007,Lietal.[94]proposedtheirmicrofluidicdeviceforsinglecellanalysis.Thechipwasfabricatedbystandard1-photomaskwithlowcostmethod.Thedeviceconsistsfourreservoirs,fourchannelsandoneopenregionwhichcontainacellretentionchamber.Reservoironewasusedforcellintroductionandwashing,whereasreservoirtwowasusedforreagentdelivery.Reservoirthreeandfourwereusedaswastereservoirs.Byusingthischip,theyquantifieddynamicCa2+mobilizationofasinglecardiomyocyteduringitsspontaneouscontraction.Also,theymonitoredsuccessfullydynamicresponsesfromvariousexternalstimulationsuchasdaunorubicin(cardiotoxicchemotherapeuticdrug),caffeine,andisoliquiritigenin(herbalanticancer).TheirresultsalsoprovethatanticancerdrugshavelesseffectontheCa2+ofthecardiomyocytes.Thisdevicehasquantifiedthecellularresponseofsinglecardiomyocytes,discoveryofheartdiseasesdrugandcardiotoxicitytesting.
In2010,Mellorsetal.[95]proposedanelectrophoreticandelectrosprayionizationbasedmicrofluidicdeviceforsinglecellanalysis.Thedevicewasfabricatedoncorningborosilicateglasssubstratebyusingstandardphotolithographyandwetchemicaletchingtechnique.Figure12showstheschematicofthemicrofluidicdevice,whereAwasacellloadingreservoirandBwasbufferloading,whichintersectswiththeseparationchannel
Thisintersectionzonewasacelllysiszone.CSwasanelectro-osmoticpumpwhichwasconnectedwithanelectrophoreticseparationchannelandelectrosprayorifice.Cellscanflowthroughhydrodynamicallyorelectricallytotheintersectionzone,wherecellswereelectricallylysed.Then,cellscanmigratetoelectrosprayorificethroughtheseparationchannelwherecellselectrosprayionizationoccurred.Thisdevicesuccessfullylysedhumanerythrocyteswithreal-timeelectrophoreticseparation.Thehemegroup,αandβsubunitsofhemoglobinweredetectedfromerythrocyteswhencellswerecontinuouslyflowedthroughthedevice.Thisdevicecananalyze12c/m.
Localizedsinglecellmembraneelectroporationcanprovidebettercelltransfectionwithmicro/nanofluidicdevicescomparedtosinglecellelectroporation(SCEP)orbulkelectroporation(BEP).Becauseofmicro/nanoscaleelectrodedimensionanddistancebetweentwoelectrodeswereverysmall,asaresult,electricfieldcanintenseinaverysmallregionofthecellmembranecomparedtosinglecelldimension.Thus,thelocalareaofthesinglecellcanbeaffectedbyastrongelectricfield,whereasotherareaswillbeunaffected.Duetotheeffectsofsmallareasofthewholesinglecell,highcellviabilityandhightransfectionratecanbeachievedcomparedtosinglecellelectroporation.However,Boukanyetal.showlocalizedsinglecellelectroporationbyusingnano-channelbasediontransportationusingelectrophoresismethodwithtwolargeelectrodes[30].Byfabricatingmicro/nanoelectrodeswithamicro/nanoscaleelectrodegap,thisdevicecanprovidesomepromisingparameterssuchaslowvoltageandpowerrequirement,lowertoxiceffectduetonegligibleiongeneration,smallsamplevolumeandnegligibleheatgeneration.Theseparametersareessentialtoachievehightransfectionrateandhighcellviability.Thus,microfluidicbasedLSCMEPprocesscanprovideabetterunderstandingtoanalyzeintracellularcytosoliccompoundscomparedtoSCEPorthebulkelectroporationprocess.Nawarathnaetal.,demonstratedtheAFMbasedLSCMEPprocess.Figure13showslocalizedelectroporationofasinglecellusingatomicforcemicroscopy(AFM)technique[29].Forthisexperiment,theymodifiedAFMtiptoactasanano-electrodetomakeanintensehighelectricfieldnearthelocalizedareaofthesinglecellmembrane.AborondopedsiliconAFMtips(σ=0.001Ωcm,k=1.5N/m)wasusedforLSCMEPprocess.Beforeelectroporation,thetipwasgrownwith20nmSiO2layerandfinallythisoxidizedtipwassectioneduntilbaresiliconwasexposedbyfocusedionbeam(FIB)technique.Asaresult,asmallerareaofbaresiliconcancauseanintensehighelectricfieldonasinglecellmembrane.Theyhavereducedthisbaresiliconareadownto0.5μmindiameter,whichwasconcentratedwithanintenseelectricfieldon10μmdiameterofratfibroblastcell.Figure13a–hshowstheresultsofLSCMEPtechniqueusingAFMtipforelectroporationprocessandFigure13idemonstratedtheAFMtip,whichwaspositionedontopofthesinglecellforlocalizedsinglecellmembraneelectroporation(LSCMEP)process.Tomakeanintensehighelectricfield,1Vppwith0.5Hzpulsewasusedtotransfectratfibroblastcells.Thetransfectionofsinglecellwascompletedwithin10s.Thisdevicecanperformhighlylocalizedelectroporationofasiglecellwithconcentricelectricfieldonlocalareaofsinglecellmembrane.Theexperimentcanbeperformedinafriendlyenvironmentsuchascellculturedishes,etc.
Inrecentyears,Boukanyetal.[30]showednanochannelbasedlocalizedsinglecellelectroporationwithapreciseamountofbiomoleculesdelivery.Inthisdevice,theypositionedasinglecellinonemicrochannelbyopticaltweezersandtransfectionagentwasloadedtoanothermicrochannel.Thesetwomicrochannelswereconnectedbyonenanochannel.Toapplyaveryhighelectricfieldinbetweentwomicrochannels,atransfectionagentwasdeliveredthroughthenanochannelusinganelectrophoreticallydrivenprocessandfinallydrugsweredeliveredinsideasinglecellthroughaverysmallareaofthecellmembrane.In2012,Chenetal.demonstratedanotherlocalizedsinglecellmembraneelectroporationusimgITOmicroelectrodebasedtransparentchip[27].Figure14showsmicrofluidiclocalizedsinglecellmembraneelectroporationdevice.TheydepositedITOfilmsonacoveredglasssubstrateandpatterenditbystandardlithographicprocesstoformasITOlines.Afterthat,athinSiO2layerwasdepositedasapassivationlayerbyplasmaenhancedchemicalvapordeposition(PECVD)technique.ThefinalITOlineswerecutbythefocusedionbeam(FIB)technique.Thegapbetweentwoelectrodeswere1μmandwidthofeachelectrodewas2μm.Whensinglecellwasstronglyattachedinbetweentwoelectrodesgap,theelectricfieldwasintensedinonlya1μmgapareaonsinglecellmembrane.Asaresult,theydemonstratedlocalizedsinglecellmembraneelectroporationwithmicrofluidicdevice.Figure14ashowslocalizedelectroporationprocessbetweentwomicro-electrodesandFigure14bshowsmultiplenumberofelectrodesforLSCMEPprocess.Figure14canddshowstheopticalmicroscopeimageofpatterenedITOmicroelectrodesandscanningelectronmicroscope(SEM)imageofITOmicroelectrodsewithmicro-channel.Accordingtotheirresults,theyachived0.93μmelectroporationregionwith60%cellviabilityfor8Vpp20mspulseapplication.Toreducethegapbetweentwoelectrodes,ahightransfectionratecanbeachivedbythistechnique.Thisdevicenotonlycontroltherecoveryofcellmembranes(reversibleelectroporation)withoutcelldamagebutalsoitprovidesclearopticalviewbyusinganinvertedmicroscope(ITObasedtransparentchip).
Recently,anotherLSCMEPbaseddevicewasproposedbyJokilaaksoetal.[77]forsinglecelllysis.Theyreportedasiliconnanowireandnanoribbonbasedbiologicalfieldeffecttransistorforsinglecellpositioningandlysismechanism.Figure15ashowsthecrosssectionalviewandelectricconnectionwithPDMSabovethedeviceandFigure15bshowsanarrayofthetransistorswithbothnanowiresandnanoribons.Topositionthesinglecellonthisdevice,theyusedprogrammablemagneticfieldformagneticmanipulationof7.9μmCOOHmodifiedCOMPELmagneticmicrosphere.Afterpositioningthesinglecell(HT-29)ontopofthetransistor,cellswereadheredfor30minpriortoelectroporationexperiment.Theappliedelectricfieldwas600–900mVpp(peaktopeak)at10MHzfor2mspulse.Thiselectricfieldwasconnectedwithashortedsourceanddraininoneterminalandanotherterminalconnectedonthegateofthedevice.Theelectricfieldintensitywasfringinginnature,whichaffectedthecellmembraneintegrityleadingtocelllysis.Thisdevicecanperformsinglecelllysiswhichispotentiallyapplicabletomedicaldiagnosticsandbiologicalcellstudies.
Insummary,thisarticledescribesthedetailsaboutbulkelectroporation(BEP),singlecellelectroporation(SCEP),andlocalizedsinglecellmembraneelectroporation(LSCMEP)byusingmicro/nanofluidicdeviceswiththeiradvantagesanddisadvantages.Alloftheseprocessescandeliverdrugs,DNA,RNA,oligonucleotides,proteins,etc.However,toanalyzecelltocellbehaviorwiththeirorganellesandintracellularbiochemicaleffect,singlecellanalysismustbeexecuted.Micro/nanofluidicdevicesarethepotentialcandidatestoanalyzesinglecells,becauseoftheirdimensionreductiontothedimensionofsinglecelllevel.Thesedevicesprovideeasyperformancesuchascellhandling,lowerpowerconsumption,lowtoxicity,smallsamplevolume,lowercontaminationrate,highcellviability,andhightransfectionratewhencomparedtoconventionalelectroporation.Toreducetheelectrodeareaandgapbetweentwoelectrodesbyusingmicro/nanofluidicdevices,selectiveandlocalizeddrugdeliveryispossible.Thisnewapproachiscalledlocalizedsinglecellmembraneelectroporation(LSCMEP).However,untilnowthistechniqueisinthedevelopmentstage.Inthefuture,theLSCMEPprocesscanprovideselectiveandspecificsinglecellmanipulationfrommillionsofpopulationsofcellstog